KR101948805B1 - Imidazo[2,1-b]oxazole derivatives with anti-tumor activity and pharmaceutical compositions comprising the same - Google Patents

Abstract

There is provided a pharmaceutical composition for the prevention and treatment of tumors which comprises as an active ingredient an imidazooxazole derivative compound, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, The tumor can be prevented or treated by administration.

Description

[0001] The present invention relates to an imidazooxazole derivative having an antitumor effect and a pharmaceutical composition comprising the same. [0002] Imidazo [2,1-b] oxazole derivatives with anti-

The present invention relates to an imidazooxazole derivative or a pharmaceutically acceptable salt thereof, a process for producing the same, and a pharmaceutical composition for preventing or treating tumor comprising the same as an active ingredient.

MAPK signaling pathway (EGFR> Ras> Raf> Mek> Erk) is a signaling pathway involved in cell proliferation.

There are three known RAF kinase enzymes in humans (Marais and Marshall Cancer Surv. 27: 101-125, 1996), A-RAF, B-RAF and C-RAF. RAF protein has been recognized as an important source and promoter of malignant tumors as mutation of B-RAF has been found to be a high frequency in human tumors (Davies, H. et al., Nature 417: 949-954 (2002)).

In the case of B-RAF, approximately 7% of the mutations are found in relation to the tumors and the frequency of melanoma (50-70%), ovarian cancer (35%), thyroid cancer (50%) and rectal cancer (Tuveson, et al., Cancer Cell. 4: 95-98 (2003); and Xing, Endocrine-Related Cancer: 12: 245-262 (2005)). Approximately 90% of these mutations are V600E in which the valine residue of the kinase domain is point mutated to glutamate, which is an important target for the development of anticancer drugs. V600E B-RAF has over 500-fold higher phosphorylation, resulting in hyperactivation of MEK, ERK and abnormal growth of tumor cells. To date, approximately 40 B-RAF mutants have been identified, most of which occur in active fragment sites and glycine-rich G-loops located in the kinase activity domain, and the incidence of other mutants other than V600E is significantly lower. About 10% of the B-RAF mutants in rectal cancers occur in the G-loop of the kinase domain (Rajagopalan et al., Nature 2002 418, 934).

Recent studies have shown that inhibition of mutant B-RAF by siRNA in human melanoma cells inhibits both MEK and ERK, resulting in arresting tumor cell growth and ultimately accelerating cell death (Sharma, et Also in the xenograft model test of B-RAF mutation using short-hairpin RNAs, it is also possible to use a short-hairpin RNA Inhibition of B-RAF has been shown to inhibit and reversibly regulate tumors (Hoeflich et al., Cancer Res. 66: 999-1006 (2006) B-RAF is an important target of anticancer drugs.

One aspect is to provide an imidazooxazole derivative compound, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention is to provide a pharmaceutical composition for preventing and treating tumors which comprises the above compound, solvate, stereoisomer or pharmaceutically acceptable salt thereof as an active ingredient.

Yet another aspect is to provide a method of preventing or treating a tumor, comprising administering the pharmaceutical composition to a subject at risk of engaging or taking the tumor.

One aspect is a compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof, represented by the following formula (S)

[화학식 S]

[Chemical Formula S]

In the above formula (S)

으로서,A is

As a result,

When X is O, R ¹ is hydrogen, halogen, C ₁ -C ₄ alkoxy, hydroxy or combinations thereof,

When X is S, R ¹ is hydrogen, Cl, Br, I, C ₁ -C ₄ alkoxy, hydroxy or combinations thereof,

또는

으로서,B is

or

As a result,

n = 1 or 2,

R ² , R ³ and R ⁴ are each independently hydrogen, tert-butyloxycarbonyl or C ₁ -C ₄ linear or branched alkyl unsubstituted or substituted with one or more substituents, C ₅ -C ₁₂ Aryl, C ₅ -C ₁₂ heteroaryl, or C ₅ -C ₁₂ heterocycloalkyl,

R ⁵ is X 'or X'-Y, wherein each of X' and Y is independently selected from the group consisting of C ₅ -C ₁₂ aryl, C ₅ -C ₁₂ heteroaryl, C ₅ -C ₁₂ Aryl C ₁ -C ₄ alkyl, C ₅ -C ₁₂ heteroaryl C ₁ -C ₄ alkyl, C ₅ -C ₁₂ cycloalkyl or C ₅ -C ₁₂ heterocycloalkyl,

Wherein the substituents are halogen, C ₁ ~ C ₄ linear or branched alkyl, C ₁ ~ C ₄ alkoxy, trifluoromethyl, hydroxy, amine, C ₁ ~ C ₄ alkyl amine, nitro, amide, C ₁ ~ C ₄ < / RTI > alkyl amide, urea and acetyl.

In one embodiment, when X is O, R ¹ is hydrogen, fluorine, methoxy, hydroxy, or combinations thereof,

R ² , R ³ and R ⁴ are each independently hydrogen, tert-butyloxycarbonyl, ethyl, methyl or benzyl,

R < ⁵ > is phenyl, unsubstituted or substituted with one or more substituents,

Wherein said substituent is selected from the group consisting of methyl, ethyl, fluorine, trifluoromethyl, hydroxy, methoxy, nitro, amide, urea and acetyl, solvates, stereoisomers or pharmaceutically acceptable salts thereof .

The 'halogen' may be F, Cl, Br, or I.

The 'alkoxy' may be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or t-butoxy.

The term "alkyl" means a straight or branched aliphatic hydrocarbon group having a specified number of carbon atoms and may be methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, or t-butyl.

The 'aryl' may be phenyl, naphthyl, anthracenyl, indanyl or biphenyl.

Heteroaryl " or " heterocycloalkyl " means a heterocyclic ring selected from the group consisting of azetidinyl, acridinyl, benzodioxolyl, benzodioxanyl, benzofuranyl, carbazolyl, cinnolinyl, dioxolanyl, pyridyl, Pyridyl, pyrimidinyl, pyrimidinyl, pyrimidinyl, pyrimidinyl, pyrimidinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrazolyl, imidazolyl, tetrahydroisoquinolinyl, pyrrolyl, Wherein the phenyl ring is optionally substituted with one or more substituents selected from the group consisting of halogen, fluorine, fluorine, fluorine, fluorine, fluorine, fluorine, fluorine, Wherein the heteroaryl is selected from the group consisting of hydrogen, fluorine, fluorine, chlorine, fluorine, fluorine, chlorine, bromine, iodine, Piperidinyl, pyrrolidinyl, 2-oxopyrrolidinyl or oxazolidinyl.

The term "cycloalkyl" refers to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl, cycloheptyl, perhydronaphthyl, adamantyl, crosslinked cyclic groups and spirobicyclic groups Can be selected

In other embodiments, the compound may be represented by any one of the following general formulas I or IV:

[화학식 Ⅰ];

[Formula I] ;

[화학식 Ⅱ];

[Formula II] ;

[화학식 Ⅲ]; 및

[Formula III] ; And

[화학식 Ⅳ].

[Formula IV].

In the above formula, X, n, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as mentioned above.

In another embodiment, the compound represented by formula (S)

Yl) pyrimidin-2-yl) amino) ethyl) -5-methyl-2-oxo- -1,2,5-thiadiazolidine 1,1-dioxide (1I);

Yl) pyrimidin-2-yl) amino) ethyl) -1H-pyrazolo [3,4-d] pyrimidin- -1,2,5-thiadiazolidine 1,1-dioxide (2I);

Yl) pyrimidin-2-yl) amino) ethyl) -1H-pyrazolo [3,4-d] pyrimidin- -1,2,5-thiadiazolidine 1,1-dioxide (3I);

Yl) pyrimidin-2-yl) amino) ethyl) -5-methyl-2-oxo- -1,2,5-thiadiazolidine 1,1-dioxide (4I);

Yl) pyrimidin-2-yl) amino) ethyl) -1H-pyrazolo [3,4-d] pyrimidin- -1, 2,5-thiadiazolidine 1,1-dioxide (5I);

Yl) pyrimidin-2-yl) amino) ethyl) -2,3-dimethyl-lH-pyrrolo [2,3- -1,2,5-thiadiazolidine 1,1-dioxide (6I);

Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydro- , 5-thiadiazolidine 1,1-dioxide (7I);

Yl) pyrimidin-2-yl) amino) ethyl) -4, 4-dihydro- - dimethyl-1,2,5-thiadiazolidine 1,1-dioxide (8I);

Yl) pyrimidin-2-yl) amino) ethyl) -6-methyl (2-oxo- -1,2,6-thiadiazinic acid 1,1-dioxide (9I);

Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydro- , 6-thiadiazole past 1,1-dioxide (10I);

Yl) pyrimidin-2-yl) amino) ethyl) -5-methyl-2-oxo- -1,2,5-thiadiazolidine 1,1-dioxide (IIII);

Yl) pyrimidin-2-yl) amino) ethyl) -1,2,3,4-tetrahydroisoquinoline; -1,2,5-thiadiazolidine 1,1-dioxide (2 II);

2-yl) amino) ethyl) - < / RTI > -1,2,5-thiadiazolidine 1,1-dioxide (3 II);

Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydroxy- , 5-thiadiazolidine 1,1-dioxide (411);

Yl) pyrimidin-2-yl) amino) ethyl) -4, 4-dihydroxyphenyl) imidazo [ - dimethyl-1,2,5-thiadiazolidine 1,1-dioxide (5 II);

Yl) pyrimidin-2-yl) amino) ethyl) -6-methyl-2-methyl- -1,2,6-thiadiaz pastl 1,1-dioxide (6 II);

Yl) pyrimidin-2-yl) amino) ethyl) -1,2,3,4-tetrahydro- -1,2,6-thiadiazin-2-carboxylate 1,1-dioxide (711);

Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydroxy- , 6-thiadiazolidin-1,1-dioxide (8 II);

Yl) amino) ethyl) benzenesulfonamide (prepared from 4-fluoro-N- (2 - ((4- (6-phenylimidazo [2,1- b] thiazol- 1 III);

2-yl) amino) ethyl) benzenesulfonamide (prepared from 4-methoxy-N- (2- 2 III);

Amino) ethyl) benzenesulfonamide (3 < RTI ID = 0.0 > III) < / RTI &);

2-yl) amino) ethyl) benzenesulfonamide (prepared as described for the preparation of N- (2 - ((4- (6- (4- fluorophenyl) imidazo [2,1- b] oxazol-5-yl) pyrimidin- 4 IV);

Yl) pyrimidin-2-yl) amino) ethyl (2-fluoro-4- ) Benzenesulfonamide (5 IV);

Yl) pyrimidin-2-yl) amino) ethyl (2-oxo-pyridin-2-yl) ) Benzenesulfonamide (6IV);

Yl) pyrimidin-2-yl) amino) ethyl) -4-iodo-2- (4- (6- (4- fluorophenyl) imidazo [ Lt; / RTI > benzenesulfonamide (7IV);

Yl) pyrimidin-2-yl) amino) ethyl) -4-methylpiperazin-1- Lt; / RTI > benzenesulfonamide (8IV);

Yl) pyrimidin-2-yl) amino) ethyl) -4-methyl (2-oxo- Benzenesulfonamide (9IV);

Yl) amino) ethyl) -4- ((4-fluorophenyl) imidazo [ Trifluoromethyl) < / RTI > benzenesulfonamide (10IV);

Yl) pyrimidin-2-yl) amino) ethyl) -3, 4-dihydro- - dimethoxybenzenesulfonamide (11IV);

Yl) pyrimidin-2-yl) amino) ethyl) -3, 4-dihydro- - dimethylbenzenesulfonamide (12IV);

Yl) pyrimidin-2-yl) amino) ethyl) naphthalene-2-carboxylic acid methyl ester was used in place of N- (2- (4- (6- (4- fluorophenyl) imidazo [ Sulfonamide (13 IV);

2-yl) amino) ethyl) benzenesulfonamide (prepared according to the procedure described for the synthesis of N- (2 - ((4- (6- (3- methoxyphenyl) imidazo [2,1- b] thiazol- 4);

Yl) pyrimidin-2-yl) amino) ethyl (2-methyl-2- ) Benzenesulfonamide (5 < / RTI >III);

Yl) pyrimidin-2-yl) amino) ethyl < / RTI > ) ≪ / RTI > benzenesulfonamide (6III);

2-yl) amino) ethyl) -4-methyl-2-oxo- Benzenesulfonamide (7III);

2-yl) amino) ethyl) -4- ((4-fluoropyrimidin- Trifluoromethyl) benzenesulfonamide (8 < / RTI >III);

Yl) pyrimidin-2-yl) amino) ethyl < / RTI > ) Benzenesulfonamide (9 < / RTI >III);

Yl) pyrimidin-2-yl) amino) ethyl) naphthalene-2-carboxylic acid methyl ester was used in place of N- (2- (4- (6- (3- methoxyphenyl) imidazo [ Sulfonamide (10 III);

Yl) pyrimidin-2-yl) amino) ethyl (2-oxo-2- ) Benzenesulfonamide (11 < / RTI >III);

Yl) pyrimidin-2-yl) amino) ethyl (2-methyl-2- ) Benzenesulfonamide (12III);

2-yl) amino) ethyl) -4-methyl-2-oxo- Benzenesulfonamide (13 < / RTI >III);

2-yl) amino) ethyl) naphthalene-2-carboxylic acid methyl ester of N- (2 - ((4- (6- (4- methoxyphenyl) imidazo [ Sulfonamide (14III);

Yl) amino) propyl) benzenesulfonamide (prepared from 4-fluoro-N- (3 - ((4- (6-phenylimidazo [2,1- b] thiazol- 15 III);

2-yl) amino) propyl) benzenesulfonamide (prepared from 4-methoxy-N- (3 - ((4- (6-phenylimidazo [2,1- b] thiazol- 16 III);

Yl) amino) propyl) benzenesulfonamide (17 < RTI ID = 0.0 > III < / RTI >);

Yl) amino) propyl) benzenesulfonamide (prepared according to the procedure described for the synthesis of N- (3 - ((4- (6- (4- fluorophenyl) imidazo [2,1- b] oxazol-5-yl) pyrimidin- 28 IV);

Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide (29 IV);

Yl) pyrimidin-2-yl) amino) propyl) -4-iodo-2- (4- (6- (4- Lt; / RTI > benzenesulfonamide (30IV);

Yl) pyrimidin-2-yl) amino) propyl) -4-methoxy-N- (3- Lt; / RTI > benzenesulfonamide (31 IV);

Yl) pyrimidin-2-yl) amino) propyl) -4-methyl-lH-pyrrolo [2,3-d] pyrimidin- Benzenesulfonamide (32IV);

Yl) amino) propyl] -4- ((4-fluorophenyl) imidazo [ Trifluoromethyl) benzenesulfonamide (33 < IV >);

Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide (34 < IV >);

Yl) pyrimidin-2-yl) amino) propyl) -3, 4-dihydro- - dimethoxybenzenesulfonamide (35IV);

Yl) pyrimidin-2-yl) amino) propyl) -3, 4-dihydro- - dimethylbenzenesulfonamide (36IV);

Yl) pyrimidin-2-yl) amino) propyl) naphthalene-2-carboxylic acid methyl ester was used in place of N- (3- (4- (6- (4- fluorophenyl) imidazo [ Sulfonamide (37IV);

2-yl) amino) propyl) benzenesulfonamide (prepared according to the procedure described for the synthesis of N- (3 - (4- (6- (3- methoxyphenyl) imidazo [2,1- b] thiazol- 18 III);

Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide (19 < / RTI >III);

Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide (20III);

Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide (21 < / RTI >III);

Yl) amino) propyl) -4-methylpyridin-2-yl) Benzenesulfonamide (22 < / RTI >III);

Yl) pyrimidin-2-yl) amino) propyl) naphthalene-2-carbaldehyde, N- (3- (4- (6- (3- methoxyphenyl) imidazo [ Sulfonamide (23III);

2-yl) amino) ethyl) benzenesulfonamide (prepared from 4-hydroxy-N- 24 III);

Yl) pyrimidin-2-yl) amino) ethyl) -4-hydroxy-2-oxo- Roxybenzenesulfonamide (45IV);

Yl) pyrimidin-2-yl) amino) ethyl < / RTI > ) Benzenesulfonamide (25 < / RTI >III);

Yl) pyrimidin-2-yl) amino) ethyl < / RTI > ) Benzenesulfonamide (26 < / RTI >III);

2-yl) amino) ethyl) -4-methyl-2-pyrrolidinone Benzenesulfonamide (27III);

Yl) pyrimidin-2-yl) amino) ethyl (2-oxo-2- ) Benzenesulfonamide (28 < / RTI >III);

Yl) amino) ethylbenzenesulfonamide (29 < RTI ID = 0.0 > (III) < / RTI >);

Yl) pyrimidin-2-yl) amino) propyl) -4-hydroxy-2, Roxybenzenesulfonamide (51 IV);

2-yl) amino) propyl) benzenesulfonamide (prepared by the method described in Example 1, step 2) and N- (3 - ((4- (6- (3- hydroxyphenyl) imidazo [2,1- b] thiazol- 30 III); And

Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide (31 < RTI ID = 0.0 > III). ≪ / RTI >

One aspect includes cyclizing Formulas (1) and (2) to produce a compound of Formula (3);

Reacting a compound of formula 3 with 4-chloro-2- (methylthio) pyrimidine to produce a compound of formula 4;

Oxidizing the compound of formula (4) to produce a compound of formula (5); And

Reacting a compound represented by the formula (5) with an N-aminoethylcyclic sulfonamide derivative or an N- (2-aminoethyl) benzenesulfonamide derivative under a base to obtain a compound represented by the formula S, a compound represented by the formula Cargo, stereoisomers, or pharmaceutically acceptable salts thereof.

Specifically, the above production method can be represented by the following Reaction Scheme 1.

[Reaction Scheme 1]

Wherein A, B, and X are as defined above.

In one embodiment, the step of reacting the compound of formula (3) with 4-chloro-2- (methylthio) pyrimidine to produce the compound of formula (4) may be carried out on the basis of a Heck reaction, Pd (OAc) ₂ ≪ / RTI > Each step may further comprise a step of purifying as needed. In step 4, the base may be diisopropylethylamine (DIPEA).

An aspect of the present invention provides a pharmaceutical composition for preventing and treating tumors, which comprises the above compound, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof as an active ingredient.

Another aspect provides the use of a composition comprising said compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof for the manufacture of said pharmaceutical composition.

The term " pharmaceutically acceptable salt " refers to a concentration that is relatively non-toxic to a patient and has a beneficial effect that is harmless, such that the side effects caused by the salt are any organic or inorganic species of the compound of formula S that does not impair the beneficial efficacy of the compound of formula Means an addition salt. Examples of the inorganic acid include hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid and phosphoric acid. Examples of the organic acid include citric acid, acetic acid, lactic acid, maleic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, maleic acid, tartaric acid, succinic acid, malonic acid, succinic acid, malonic acid, glutamic acid, aspartic acid, oxalic acid, P-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid. These salts also include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, the acid addition salt may be selected from the group consisting of acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camylate, citrate, eddylate, Hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, malate, glucoside, gluconate, gluconate, glucuronate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / Hydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydroxyacetate, Lactate, stearate, succinate, tartrate, tosylate, trifluoroacetate Diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salts, and the like. Lt; / RTI > hydrochloride or trifluoroacetate.

In one embodiment, the compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof may inhibit protein kinase and inhibit tumor cell proliferation. The protein kinase is involved in tumor cell proliferation and is involved in the proliferation of tumor cells such as V600E RAF (Rapidly Accelerated Fibrosarcoma), B-RAF, C-RAF, Mitogen-activated protein kinases 14 (MAPK14), FLT3 (Fms-like tyrosine kinase 3) synthase kinase 3 beta).

In another embodiment, the tumor is selected from the group consisting of lung cancer, liver cancer, esophageal cancer, gastric cancer, small bowel cancer, small bowel cancer, pancreatic cancer, melanoma, breast cancer, oral cancer, bone cancer, brain tumor, thyroid cancer, Cancers such as urothelial carcinoma, bladder cancer, testicular cancer, leukemia, multiple myeloma, myelodysplastic syndrome, or hematologic cancer such as glioblastoma, Hodgkin's disease or non-Hodgkin's lymphoma, Behcet's disease, skin cancer, psoriasis and fibrosarcoma.

One aspect provides a method of preventing or treating a tumor comprising administering the compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof to a subject at risk of acquiring or engaging with the tumor.

The subject may be a mammal, and the mammal may be a human, and the effective dosage of the compound of the present invention on the human body may vary depending on the age, weight, sex, dosage form, have.

Such administration can be carried out in various formulations for oral administration or for parenteral administration such as intravenous, intraperitoneal, subcutaneous, subcutaneous, epithelial or muscular administration. In the case of formulation, the fillers, extenders, binders, wetting agents, Release agents, surfactants, and the like.

Solid formulations for oral administration may include tablets, patients, powders, granules, capsules, troches and the like, which may contain one or more excipients such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. As the liquid preparation for oral administration, suspensions, solutions, emulsions or syrups may be used. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, .

Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

FIG. 1 shows the anticancer effect after administration of Compound 2II in the xenograft model of A375P cell line in terms of tumor size according to days after transplantation. G1 represents the vehicle control group, G2 represents the test substance 20 mg / kg / day administration group, and G3 represents the test substance 50 mg / kg / day administration group.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are intended to illustrate the present invention, and the scope of the present invention is not limited by these examples.

Production Method 1

Compounds of formulas I-IV were prepared according to the following procedure as shown in Schemes 2 and 3 below.

The synthesis of the imidazooxazole compound is as shown in the following Reaction Scheme 2. In order to synthesize the compounds of Formulas I to IV, the core intermediate compound 5 It is important to synthesize.

Scheme 2

2-Aminooxazole (2) was cyclized by reacting with? -Bromo-4-fluoroacetophenone and ethanol under reflux for 16 hours, and compound 3 was synthesized through crystallization using ammonia water. Compound 3 thus synthesized was reacted with 4-chloro-2- (methylthio) pyrimidine at 80 ° C for 36 hours under a Pd (OAc) ₂ catalyst based on Heck reaction conditions. After completion of the reaction, methylthiopyrimidinyl compound 4 was obtained by column chromatography. Compound 4 thus synthesized was oxidized with oxone at room temperature through an over-night reaction and purified by column to synthesize Compound 5 as a core intermediate. Aminoethylcyclic sulfonamide or N- (2-aminoethyl) benzenesulfonamide is dissolved in DMF solvent under diisopropylethylamine and stirred at 80 占 폚 for 5 to 10 hours under diisopropylethylamine to give Compound I, II And IV. The double methoxy compound was dissolved in methylene chloride, and the mixture was cooled to -78 ° C. Then, BBr ₃ was slowly added dropwise and stirred for 30 minutes. The temperature was elevated to room temperature and demethylated for 12 hours to synthesize the desired compound having a hydroxyl group.

Production Method 2

The synthesis of the imidazolothiazole compound was carried out in the same manner as in Reaction Scheme 2 except that 2-aminothiazole (2) was used in the same manner as in Reaction Scheme 3 below.

Scheme 3

The compound of each step is shown in the following examples.

Example  1. 6- Phenylimidazo [2,1-b] oxazole  (3a in scheme 2)

2-Aminooxazole ( 2 , 4.7 g, 47.4 mmol in Scheme 2) and? -Bromo-4-acetophenone (10.8 g, 49.9 mmol) were added to acetonitrile (120 mL) and stirred at room temperature for 48 hours . The reaction was terminated by TLC and then filtered. The resulting solid was taken up in toluene (100 mL). To the suspension was added slowly 20 mL of titanium chloride (1M dichloromethane solution) at 0 < 0 > C. The reaction solution is refluxed and stirred for 20 hours. The solution was concentrated to half its original volume. Add saturated sodium carbonate solution and extract with ethyl acetate. The organic layer was dried, filtered using anhydrous Na ₂ SO _4. Distilled under reduced pressure and concentrated to obtain the desired compound 6- (4-phenyl) imidazo [2,1- b ] oxazole (3a) in crystalline form.

White solid (5.1 g, 56%). mp 143-144 [deg.] C; ^{IR (KBr) [cm -1]} : 3139, 3120, 1955, 1884, 1602. 1 H-NMR (DMSO- d 6, 300 MHz) δ 8.63 (d, 1H, J = 7.17 Hz), 8.29 (t, 1H, J = 5.71 Hz), 7.77 (t, 3H, J = 6.7 Hz), 7.49-7.36 (m, 3H). _{^{13 CNMR (DMSO- d 6, 75}} MHz) δ 158.1, 154.4, 134.9, 132.6, 127.7, 125.7, 121.9, 119.6, 111.5, 104.2.

Example  2. 5- (2- ( Methyl thio ) Pyrimidin-4-yl) -6- Phenylimidazo [2,1-b] oxazole  (4a of Scheme 2)

6- (4-phenyl) imidazo [2,1- b] oxazole (the compound 3a, 8.26 g, 37.8 mmol) , 4- chloro-2- (methylthio) pyrimidine (9.09 g, 56.6 mmol), Cesium carbonate (18.44 g, 56.6 mmol), palladium acetate (1.71 g, 7.6 mmol) and triphenylphosphine (3.96 g, 15.1 mmol) were added to DMF (80 mL) and stirred at 80 ° C for 36 hours. After completion of the reaction was confirmed by TLC, the reaction mixture was cooled to room temperature, EtOAc (200 mL) and distilled water (250 mL) were added, and the layers were separated. The aqueous layer was extracted three times with EtOAc (100 mL) and the aqueous layer was discarded. The organic layer was washed with saline (100 mL), dried over anhydrous Na ₂ SO ₄ and filtered. The residue was purified by flash column chromatography to obtain the desired compound, 6- (4-phenyl) -5- (2- (methylthio) pyrimidin-4-yl) b ] oxazole ( 4a ).

White solid (19%), mp. 143-144 DEG C; IR (KBr) [cm- ¹ ]: 3054, 6103, 1538, 1685; _{^{1 H-NMR (CDCl 3,}} 300 MHz) δ 8.62 (dd, 1H, J = 4.53, J = 0.7), 8.23 (dd, 1H, J = 5.4, J = 0.7), 7.65-7.62 (m, 2H) , 7.47-7.45 (m, 3H), 6.97 (dd, 1H, J = 4.53, J = 0.66), 6.89 (dd, 1H, J = 4.77, J = 0.66), 2.64 (s, 3H); ¹³ C NMR (CDCl ₃ , 75 MHz) 隆 172.4, 156.2, 156.2, 152.7, 151.1, 134.6, 129.2, 128.9, 128.8, 122.2, 120.1, 112.7, 112.0, 14.2.

Example  3. 5- (2- ( Methylsulfonyl ) Pyrimidin-4-yl) -6- Phenylimidazo [2,1-b] oxazole e  (5a of Scheme 2)

6- (4-phenyl) -5- (2- (methylthio) pyrimidin-4-yl) imidazo [2,1- b] oxazole (the compound 4a, 3.0 g, 8.8 mmol) to MeOH (300 mL), oxone (14.76 g, 97.0 mmol) in distilled water (60 mL) was added while stirring at room temperature, and the mixture was stirred for 16 hours. After completion of the reaction was confirmed by TLC, the organic solvent was removed by distillation under reduced pressure. DCM (60 mL) was added to the aqueous layer and the layers were separated. The aqueous layer was extracted three times with DCM (30 mL) and the aqueous layer was discarded. The organic layer was washed with brine (50 mL) and distilled water (50 mL), dried over anhydrous Na ₂ SO ₄ , and filtered. Concentrated by evaporation under reduced pressure and purified by flash column chromatography to obtain the desired compound 6- (4-phenyl) -5- (2- (methylsulfonyl) pyrimidin-4-yl) imidazo [ -b ] oxazole ( 5a ).

White solid (85%); _{^{1 H-NMR (CDCl 3,}} 300 MHz) δ 8.89 (d, 1H, J = 4.30), 8.56 (d, 1H, J = 5.40), 7.71 (d, 2H, J = 2.50), 7.64-7.45 (m , 3H), 7.42 (d, IH, J = 5.20), 7.17 (d, IH, J = 4.40), 3.45 (s, 3H); ¹³ C NMR (CDCl ₃ , 75 MHz) 隆 165.7, 157.5, 156.8, 153.3, 134.1, 133.3, 130.0, 129.5, 129.1, 129.0, 128.3, 119.5, 117.5, 113.8, 39.3.

Example  4. 6- (4- Fluorophenyl ) -5- (2- (methylsulfonyl) pyrimidin-4-yl) imidazole [2,1-b] Oxazole  (5c of Scheme 2)

Was synthesized by an experimental method similar to that of Compound 5a .

Yellow solid (85%); mp. 115-116 [deg.] C. _{^{1 H-NMR (CDCl 3,}} 300MHz) δ 8.90 (d, 1H, J = 4.5 Hz), 8.53 (d, 1H, J = 5.6 Hz) 7.63 (q, 2H, J = 4.7 Hz), 7.33 (d, 1H, J = 5.6 Hz), 7.22 (t , 2H, J = 8.6 Hz), 7.10 (d, 1H, J = 4.5 Hz), 3.37 (s, 3H); ^{13 C} NMR (CDCl ₃ , 75 MHz)? 171.1, 165.7, 157.3, 157.0, 154.2, 151.7, 131.0, 130.9, 129.9, 123.1, 119.4, 117.4, 116.4, 116.1, 114.2, 39.2.

Example  5. 5- (2- ( Methylsulfonyl ) Pyrimidin-4-yl) -6- Phenylimidazo [2,1-b] thiazole  (5d in scheme 3)

Was synthesized by an experimental method similar to that of Compound 5a .

White solid (95%); mp. 176-177 [deg.] C. IR (KBr) [cm- ¹ ]: 3137, 3110, 2931, 1979, 1902, 1797, 1444, 1373; _{^{1 H-NMR (CDCl 3,}} 300 MHz) δ 8.99 (d, 1H, J = 4.41), 8.58 (d, 1H, J = 5.40), 7.73 (d, 2H, J = 2.46), 7.66-7.47 (m , 3H), 7.45 (d, IH, J = 5.22), 7.17 (d, IH, J = 4.41), 3.47 (s, 3H); _{^{13 CNMR (CDCl 3, 75 MHz}} ) δ 165.7, 157.5, 156.8, 153.3, 134.1, 133.3, 130.0, 129.5, 129.1, 129.0, 128.3, 119.5, 117.5, 113.8, 39.3; LC-MS: predicted value calculated by C ₁₆ H ₁₂ N ₄ O ₂ S ₂ m / z: 356. Found: 357 (M + 1) ⁺

Example  6. 2- methyl -1,2,5- Thiadiazolidine  1,1- Dioxid  (6a)

N-Methylethyldiamine (1.5 g, 20.8 mmol) and sulfurodiimide (2.0 g, 20.8 mmol) were added to a pyridine solvent (20 mL) and heated to reflux with stirring for 3 hours. After the reaction was terminated by extraction with ethyl acetate and then the reaction was concentrated was added to toluene (5 mL), and purified by column chromatography (EtOAc) of the residue to remove the dried, filtered and the solvent separated organic layer over MgSO ₄ Compound 6a .

White solid (52.3%). Mp 80 to 82 占 폚; _{^{1 H-NMR (CDCl 3,}} 300MHz) δ7.27 (s, 1H), 3.53-3.19 (m, 2H), 3.42-3.37 (m, 2H), 2.75 (s, 1H); _{^{13 C NMR (CDCl 3, 75}} MHz) δ 32.67, 39.64, 46.94; LC-MS: calculated calculated value for C ₃ H ₈ N ₂ O ₂ S m / z: 136 found: 137 (M + 1) ⁺ .

Example  7. benzyl  (2- (5- methyl -1,1- Dioxido -1,2,5- Thiadiazolidine Yl) ethyl) carbamate (7a)

Compound 6a (0.856 g, 6.26 mmol) was dissolved in 5 mL of anhydrous DMSO under nitrogen atmosphere, and then 60% NaH (0.282 g, 11.8 mmol) was slowly added dropwise at 0 ° C. The mixture was heated to room temperature and stirred for 1 hour. 2 - (((benzyloxy) carbonyl) amino) ethyl methanesulfonate (1.5 g, 5.83 mmol) was slowly added dropwise at 0 ° C and the mixture was stirred at room temperature for 5 hours. After completion of the reaction, the reaction mixture was extracted with ethyl acetate. The organic solvent layer was dried over MgSO ₄ , filtered to remove the solvent, and the residue was purified by column chromatography (EtOAc) to obtain Compound 7a (0.82 g).

Viscous oil (42.9%). _{^{1 H-NMR (CDCl 3,}} 300MHz) δ7.36-7.30 (m, 5H), 5.19 (bs, 1H), 5.10 (s, 2H), 3.46 (q, 2H, J = 5.9 Hz), 3.36-3.26 (m, 4H), 3.20 (t, 2H, J = 6.0 Hz). LC-MS: calculated predicted value for C ₁₃ H ₁₉ N ₃ O ₄ S m / z: 313, found: 314 (M + 1) ⁺ .

Example  8. Synthesis of 2- (2- Aminoethyl ) -5- methyl -1,2,5- Thiadiazolidine  1,1-dioxide (8a)

The compound 7a (0.82 g, 2.5 mmol) was dissolved in 10 mL of methanol, and 10% Pd / C (0.40 g) was added thereto. The mixture was reacted for 2 hours by hydrogenation and filtered to remove the solvent by distillation under reduced pressure. It was used in the next reaction without purification (yield: 91%).

Example  9. N- (2- Aminoethyl )-4- Fluorobenzenesulfonamide  (11b)

White solid (72%); mp. 108 to 109; IR (KBr) cm ^{- 1} : 3582, 3350, 3299, 3107, 3068, 1903, 1317, 838; _{^{1 H-NMR (CDCl 3,}} 300 MHz) δ7.84-7.79 (m, 2H), 7.11 (t, 2H, J = 6 Hz), 2.89 (t, 2H, J = 6 Hz), 2.71 (t, 2H, J = 6 Hz). ¹³ C NMR (CDCl ₃ , 75 MHz)? 166.7, 136.0, 129.7 ( J = 33), 116.3 ( J = 90), 45.2, 40.9. LC-MS: calculated predicted value for C ₈ H ₁₁ FN ₂ O ₂ S m / z: 218 Found: 219 (M + 1) ⁺ .

Example  10. 2- (2 - ((4- (6- (4-fluorophenyl) imidazo [2,1-b] oxazole Yl) pyrimidin-2-yl) amino) ethyl) -5- methyl -1,2,5- Thiadiazolidine  1,1- Dioxid  (1I)

Light yellow solid; To a solution of compound 6- (4-fluorophenyl) -5- (2- (methylsulfonyl) pyrimidin-4-yl) imidazo [2,1- b ] thiazole (Compound 5c , 0.19 (compound 8a , 0.27 g, 1.24 mmol), DIPEA (0.29 mL, 0.20 mmol) and 2- (2-aminoethyl) -5- , 1.70 mmol), and the mixture was stirred at 80 占 폚 for 8 hours. The mixture was cooled to room temperature and distilled water (10 mL) and EtOAc (10 mL) were added to separate the layers. The aqueous layer was extracted twice with EtOAc (10 mL) and discarded. The organic layer was washed with brine (10 mL). To the organic layer with anhydrous Na ₂ SO _4, filtered, and dried. The mixture was concentrated under reduced pressure and purified by recrystallization using DCM (1 mL) and hexane (3 mL) to obtain 0.18 g of the objective compound 1I in crystalline form.

(72%); _{^{1 H-NMR (CDCl 3,}} 300MHz) δ 8.52 (d, 1H, J = 4.5 Hz), 8.08 (s, 1H), 7.61 (q, 2H, J = 4.6 Hz), 7.13 (t, 2H, J = 8.5 Hz), 6.91 (d, 1H, J = 4.5 Hz), 6.55 (d, 1H, J = 5.5 Hz), 5.81 (bs, 1H), 3.78 (q, 2H, J = 6.1 Hz), 3.43-3.30 (m, 6 H), 2.79 (s, 3 H);

Example  11. 2- (2 - ((4- (6- (3-methoxyphenyl) imidazo [2,1-b] oxazole Yl) pyrimidin-2-yl) amino) ethyl) -5- methyl -1,2,5- Thiadiazolidine  1,1- Dioxid  (4I)

Was synthesized by an experimental method similar to that of Compound (I).

White solid (50.5%); mp 144-145 [deg.] C; _{^{1 HNMR (CDCl 3, 300 MHz}} ) 8.58 (d, 1H, J = 4.5 Hz), 8.06 (d, 1H, J = 5.3 Hz), 7.33 (t, 1H, J = 7.8 Hz), 7.23-7.19 (m , 2H), 6.98-6.90 (m, 2H), 6.58 (d, 1H, J = 5.4 Hz), 3.83 (s, 3H), 3.78 (q, 2H, J = 6.1 Hz), 3.43-3.30 (m, 6H), 2.78 (s, 3H).

Example 12. 2 - (2 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] oxazole Yl) pyrimidin-2-yl) amino) ethyl) -5- methyl -1,2,5- Thiadiazolidine  1,1- Dioxid  (1 II)

Under nitrogen, compound 4I (17.5 mg, 0.1 mmol) was dissolved in 3 mL of MC and BBr ₃ (0.02 mL of a 1M solution in MC, 0.3 mmol) was added dropwise at -78 ° C. And the mixture was stirred at the same temperature for 30 minutes. And further stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was dissolved in ethyl acetate and washed with NaHCO ₃ and H ₂ O. The organic solvent was dried over MgSO ₄ and purified by column chromatography (EtOAc: hexane = 1: 2) to obtain Compound 1II .

White solid (78.1%); mp 151-152 [deg.] C; _{^{1 HNMR (DMSO- d 6, 300}} MHz) δ 9.56 (s, 1H), 8.12 (d, 1H, J = 5.3 Hz), 7.56-7.45 (m, 2H), 7.25 (t, 1H, J = 7.9 Hz ), 7.00 (s, 1H) , 6.97 (d, 2H, J = 1.5 Hz), 6.84-6.80 (m, 1H), 6.41 (d, 1H, J = 5.3 Hz), 3.71-3.62 (m, 2H) , 3.48-3.23 (m, 4H), 3.16 (t, 2H, J = 6.0 Hz), 2.60 (s, 3H).

Example  13. 2-Ethyl-5- (2 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] oxazole Yl) pyrimidin-2-yl) amino) ethyl) -1,2,5- Thiadiazolidine  1,1- Dioxid  (2 II)

Was synthesized by the same experimental method as Compound III .

White solid (66.5%); mp 149-150 [deg.] C; _{^{1 HNMR (DMSO- d 6, 300}} MHz) δ 9.58 (s, 1H), 8.11 (d, 1H, J = 5.4 Hz), 7.56-7.45 (m, 2H), 7.25 (t, 1H, J = 8.0 Hz ), 6.99-6.80 (m, 3H), 6.41 (d, IH, J = 5.3 Hz), 3.55-3.49 (m, 2H), 3.44-3.23 (m, 4H), 3.19-3.15 2.95 (q, 2H, J = 7.2 Hz).

Example  14. 2- (2 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] oxazole Yl) pyrimidin-2-yl) amino) ethyl) -4,4-dimethyl- Thiadiazolidine  1,1- Dioxid  (5 II)

Was synthesized by the same experimental method as Compound III .

White solid (25.0%); mp 196-197 [deg.] C; _{^{1 HNMR (CDCl 3, 400 MHz}} ) δ 8.55 (d, 1H, J = 5.4 Hz), 8.04 (d, 1H, J = 5.4 Hz), 7.30-7.28 (m, 1H), 7.15-7.13 (m, 2H ), 6.95 (d, H, J = 8.1 Hz), 3.76 (t, 2H, J = 6.1 Hz), 3.33 (t, 2H, J = 5.92 Hz), 3.25 (s, 2H), 1.45 (s, 6H ).

Example  15. 2- (2 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] oxazole Yl) pyrimidin-2-yl) amino) ethyl) -6- methyl -1,2,6- Tiadia Jin  1,1- Dioxid  (6 II)

Was synthesized by the same experimental method as Compound III .

White solid (71.7%); mp 184-185 [deg.] C; _{^{1 HNMR (CDCl 3, 400 MHz}} ) δ 8.57 (d, 1H, J = 4.5 Hz), 8.01 (d, 1H, J = 5.4 Hz), 7.28-7.26 (m, 1H), 7.15-7.11 (m, 2H ), 6.88 (d, H, J = 7.7 Hz), 6.59 (d, 1H, J = 5.3 Hz), 3.67 (t, 2H, J = 5.4 Hz), 3.48 (t, 2H, J = 5.2Hz), 3.37 (t, 2H, J = 6.0 Hz), 2.75 (s, 3H), 1.78 (m, 2H).

Example  16. A composition comprising 2- (2 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] oxazole Yl) pyrimidin-2-yl) amino) ethyl) -1,2,6- Tiadia Jin  1,1- Dioxid  (8 II)

Was synthesized by the same experimental method as Compound III .

White solid (85.3%); mp 183-184 [deg.] C; _{^{1 H-NMR (CDCl 3,}} 400 MHz) δ 8.85 (s, 1H), 8.14 (d, 1H, J = 5.9 Hz), 7.78 (d, 1H, J = 4.1 Hz), 7.47 (t, 1H, J = 7.7 Hz), 7.17-7.09 (m, 3H), 6.74 (d, H, J = 6.7 Hz), 3.88 (m, 2H), 3.60 (t, 2H, J = 7.7 Hz), 3.44-3.41 , ≪ / RTI > 4H), 1.78 (m, 2H).

Example  17. 4- Fluoro -N- (2 - ((4- (6- Phenylimidazo [2,1-b] thiazole Yl) pyrimidin-2-yl) amino) ethyl) Benzenesulfonamide  (III)

Compound in a nitrogen environment 5d (327 mg, 0.92 mmol) , N- (3- amino-ethyl) -4-fluoro-benzenesulfonamide (11b, 540 mg, 2.48 mmol ), diisopropylethylamine (0.57 mL 3.3 mmol) Was added to DMSO solvent (10 mL), and the mixture was stirred at 80 ° C for 8 hours. After the reaction was completed, the reaction mixture was extracted with ethyl acetate. The organic solvent layer was dried over MgSO ₄ , filtered to remove the solvent, and the residue was purified by column chromatography (EtOAc) to obtain Compound III .

Orange solid (52.1%); mp. 121-122 DEG C; ^{1 HNMR (DMSO- d6, 300 MHz} ) δ 8.83 (bs, NH), 8.05 (d, 1H, J = 6 Hz), 7.80 (dd, 2H, J = 6, J = 3 Hz), 7.58 (dd, 4H, J = 6, J = 3 Hz), 7.46 (d, 5H, J = 6 Hz), 6.31 (d, 1H, J = 6 Hz), 2.96 (t, 2H, J = 6 Hz), 2.50 ( s, 2H); ¹³ C-NMR (DMSO- d6 , 75 MHz) δ 164.2, 162.5, 160.9, 158.2, 156.2, 151.6, 148.0, 140.9, 132.7, 131.7, 129.6, 126.8, 121.0, 115.9, 114.5, 105.7, 38.7, 29.5; LC-MS: predicted value calculated by C ₂₃ H ₁₉ FN ₆ O ₂ S ₂ m / z, 494 found: 495 (M + 1) ⁺ .

Example  18. 4- Methoxy -N- (2 - ((4- (6- Phenylimidazo [2,1-b] thiazole Yl) pyrimidin-2-yl) amino) ethyl) Benzenesulfonamide  (2III)

The compound was synthesized by the same method as that of Compound III .

White solid (65%); mp. 199-200 C; ^{1 H-NMR (MeOD, 400} MHz) δ 9-10 (bs, 2NH protons), 8.87 (d, 1H, J = 4.8 Hz), 8.50 (d, 2H, J = 7.5 Hz), 8.40 (s, 2H ), 8.28 (s, 5H), 8.16 (d, 2H, J = 7.2 Hz), 7.12 (s, 1H), 3.76 (s, 3H), 3.33 (s, 2H), 3.15 _{^{13 C NMR (DMSO- d 6,}} 100 MHz) δ 163.8, 162.5, 161.4, 158.1, 156.2, 151.6, 148.1, 142.9, 138.0, 131.7, 130.1, 127.0, 121.0, 115.9, 114.4, 105.7, 55.3, 41.7, 29.5 . LC-MS: predicted value calculated by C ₂₄ H ₂₂ N ₆ O ₂ S ₂ m / z: 506 Found: 507 (M + 1) ⁺ .

Example  19. N- (2 - ((4- (6- (3-methoxyphenyl) imidazo [2,1-b] thiazole Yl) pyrimidin-2-yl) amino) ethyl) -4- ( Trifluoromethyl ) Benzenesulfonamide  (8 III)

The compound was synthesized by the same method as that of Compound III .

Colorless viscous oil (25%); ¹ HNMR (DMSO- d ₆ , 400 MHz)? 8.05 (d, 1H, J = 5.2 Hz), 7.73 (t, 2H, J = 7.2 Hz), 7.59 (dd, 2H, J = 2, J = 8 Hz), 7.47 (d, 4H, J = 7.2 Hz ), 7.06 (d, 2H, J = 8.8 Hz), 6.31 (d, 1H, J = 5.2 Hz), 3.79 (s, 3H), 3.39 (bs, 2H), 2.94 (brs, 2H). LC-MS: predicted value calculated by C ₂₅ H ₂₁ F ₃ N ₆ O ₃ S ₂ m / z: 574. found: 575 (M + 1) ⁺ .

Example  20. 4- Fluoro -N- (2 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] thiazole Yl) pyrimidin-2-yl) amino) ethyl) Benzenesulfonamide  (25 III)

Was synthesized by the same experimental method as Compound III .

Dark orange solid (44%); mp 70-71 [deg.] C; IR (KBr) [cm- ¹ ]: 3115, 2854, 1639, 1574, 1445, 1328; ^{1 H-NMR (MeOD, 300} ) δ 8.68 (s, 1H), 7.87 (t, 3H, J = 9 Hz), 7.58-7.45 (m, 6 H), 6.97 (d, 3H, J = 6 Hz) , 6.65 (d, 1H, J = 9 Hz), 3.33 (brs, 2H), 3.22 (t, 2H, J = 6). LC-MS: predicted value calculated by C ₂₃ H ₂₀ N ₆ O ₃ S ₂ m / z: 492. Found: 493 (M + 1) ⁺ .

Example  21. 4- Fluoro -N- (3 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] thiazole Yl) pyrimidin-2-yl) amino) propyl) benzenesulfonamide (31 III)

Was synthesized by the same experimental method as Compound III .

Light yellow solid (20%); mp 92-93 [deg.] C; IR (KBr) [cm- ¹ ]: 3382, 3258, 3118, 2933, 1731, 1574, 1447, 1331; _{^{1 H-NMR (DMSO- d 6}} , 400) δ 9.59 (s, 1H), 8.08 (d, 1H, J = 4 Hz), 7.85 (dd, 2H, J = 4, J = 8 Hz), 7.70 ( t, 1H, J = 4 Hz ), 7.45 (d, 1H, J = 4 Hz), 7.40 (t, 3H, J = 8 Hz), 7.27 (t, 1H, J = 8 Hz), 6.98 (d, 1H, J = 4 Hz), 6.84 (d, 1H, J = 8 Hz), 6.38 (d, 1H, J = 4 Hz), 3.31 (d, 2H, J = 4 Hz), 2.85 (q, 2H, J = 4 Hz), 1.70 (t, 2H, J = 4 Hz). LC-MS: predicted value calculated by C ₂₄ H ₂₁ FN ₆ O ₃ S ₂ m / z: 524. Found: 525 (M + 1) ⁺ .

< Experimental Example  1> Measurement of melanoma cell line proliferation inhibitory activity

In order to measure the cancer cell proliferation inhibitory activity of the imidazooxazole-based or imidazolothiazole-based compound according to the present invention in the cell stage, the following experiment was conducted.

(1) Screening of cancer cell lines

According to the standard protocol of the National Cancer Institute (NCI) (www.dtp.nci.nih.gov) (http://dtp.nci.nih.gov/dtpstandard/dwindex/index.jsp) Screening was performed. Briefly, human cell lines were grown in RPMI 1640 medium containing 5% FBS and 2 mM L-glutamine. For a typical screening experiment, cells are inoculated at 100 [mu] l into 96-well microtiter plates to plate at a concentration ranging from 5000 to 40,000 cells / well according to doubling time of the individual cell lines. After cell inoculation, and in the addition of the test compound to the microtiter plate before _{37 ℃, 5% CO 2,} 95% air and 100% relative humidity for 24 hours of incubation. Twenty-four hours later, two plates of each cell line were fixed in situ with trichloroacetic acid (TCA) to determine the cell population for each cell line at the time of compound addition (Tz). The test compound was dissolved in dimethyl sulfoxide at 400 times the intended final maximum test concentration and stored frozen before use. Upon compound addition, aliquots of the frozen stock were thawed and diluted to twice the final intended maximum test concentration with complete medium containing 50 μg / ml gentamicin. The control is added and the four are diluted in series of 10-fold or 1/2 log to provide five compound concentrations. An aliquot of 100 [mu] l of this different compound diluent was added to a suitable microtiter well containing 100 [mu] l of the medium already to produce the required final compound concentration.

After addition of the compound, the plate was further incubated at 37 DEG C, 5% CO2, 95% air, 100% relative humidity for 48 hours. For adherent cells, the assay was terminated by the addition of cold TCA. Cells were fixed in this state with gentle addition of 50% cold 50% (w / v) TCA (final concentration 10% TCA) and incubated at 4 ° C for 60 minutes. The supernatant was discarded and the plate was washed five times with tap water and air dried. Sulfolodamine B (SRB) solution (100 μl) at 0.4% (w / v) in 1% acetic acid was added to each well and the plate was incubated at room temperature for 10 minutes. After dyeing, the unbound dye was removed by washing with 5% acetic acid and the plate was air dried. The bound dye was solubilized sequentially with 10 mM Trizma base, and the absorbance was read with an automated plate reader at a wavelength of 515 nm. For the suspended cells, the method is the same except that the assay is terminated by fixing the precipitated cells to the bottom of the well by gentle addition of 50 [mu] l of 80% TCA (final concentration 16% TCA). Growth percentages were calculated at each compound concentration level using seven absorbance measurements (zero time (Tz), control growth rate (C) and test growth rate (Ti) in the presence of compound at five concentration levels). Growth inhibition percentages were calculated as follows.

[(Ti-Tz) / (C-Tz)] x 100 &lt; / RTI &gt;

[(Ti-Tz) / Tz] x 100 with respect to the concentration for Ti < Tz.

Table 1. IC ₅₀ values (μM) of major compounds for melanoma cell lines in vitro

In most of the melanoma cell lines shown in Table 1 above, the test compound was shown to have a proliferation inhibitory activity (all 10 μM or less), and it was confirmed that the imidazooxazole compound or imidazothiazole compound according to the present invention inhibited It can be seen that there is an excellent suppressing effect.

(2) A375P  Comparison of melanoma cell line proliferation inhibitory activity

Compared with the imidazothiazole-based compounds previously disclosed by the present inventors (MS Abdel-Maksoud et al. / European Journal of Medicinal Chemistry 95 (2015) 453-463), the compounds (2II, 9II, 15II) was more remarkably effective, the A375P cell line, a melanoma cell line, was used to compare the proliferation inhibitory activity as described below.

The A375P cell line purchased from ATCC was cultured in DMEM medium [containing 10% FBS, 1% penicillin / streptomycin] in the presence of 5% CO ₂ at 37 ° C. The cultured A375P cell line was taken with 0.05% trypsin-0.02% EDTA and 5 x 10 ³ cells per well were inoculated into a 96-well plate. To measure cell viability, MTT [3 - (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] active detection method (CellTiter 96 Assay, Promega) was used. MTT analysis followed the instructions of CellTiter 96® (Promega). The values were read at 590 nm using EnVision 2103 and the IC ₅₀ was calculated using software from GraphPad Prism 4.0.

Table 2. A375P cell line proliferation inhibitory activity of major imidazothiazole compounds IC ₅₀ value (μM)

 As shown in Table 2, it can be seen that the compounds 2II, 9II, 10II and 12II to 15II of the present invention have overall better proliferation inhibitory effect than the existing imidazothiazole compounds P1 to P5, Compared with similar structures (compounds P1 / 9II, P2 / 10II, P3 / 2II and P4 / 12II), it can be seen more clearly that the compounds of the present invention have more pronounced effects.

< Experimental Example  2> Protein Kinase  Enzyme activity measurement

The imidazooxazole-based and imidazolothiazole-based compounds according to the present invention inhibited the kinase enzyme activity in relation to the proliferation inhibitory activity of the melanoma cell line.

IC _{50 was} measured using the Reaction Biology Kinase Hotspot Service (http://www.reactionbiology.com). 25 mM Tris pH 7.5, 0.02 mM EGTA, 0.66 mg / ml myelin basic protein, 10 mM magnesium acetate and [ ³³ P-ATP] (specific activity about 500 CPM / &Lt; / RTI &gt; concentration) was incubated with kinase (5 to 10 mU). The reaction is initiated by adding a Mg-ATP mix. After incubation at room temperature for 40 minutes, the reaction was stopped by adding 5 [mu] l of 3% phosphoric acid solution. 10 μl of the reaction was spotted on a P30 Filtermat, washed once with 75 mM phosphoric acid, 5 times for 3 minutes, with methanol, and then scintillation counted.

Table 3. Effect of inhibition of kinase activity on B-RAF, V600E-RAF and C-RAF (nM)

As shown in Table 3, compounds 1II, 2II, 25III and 31III in the compounds according to the present invention showed excellent inhibitory effect on enzyme activity against B-RAF, V600E-RAF and C-RAF, Which is superior to bemulfenimb.

In summary, the compounds according to the present invention show excellent inhibitory activity against protein kinases, and in particular, various protein kinases which cause abnormal cell growth diseases such as tumor cells, such as V600E RAF, B-RAF, C-RAF , MAPK14, FLT3, and GSK3 [beta], it can be used for the prevention and treatment of tumor cell growth diseases.

< Experimental Example  3> Tumor suppression effect in melanoma mouse model

The following experiments were carried out in order to examine whether the inhibitory effect on melanoma cell proliferation and the inhibitory effect on enzyme activity on B-RAF, V600E-RAF and C-RAF were reproduced in animal models.

In this test, a xenograft model was prepared by subcutaneously administering A375P cell line, a human malignant melanoma cell line, to nude mice, and then the test substance (Compound 2II) was administered to evaluate the anticancer effect. (G1: excipient control group, G2: test substance 20 mg / kg / day administration group, G3: test substance 50 mg / kg /

The results are shown in Fig. 1 and Table 3 below.

Table 4. Increase of tumor size in melanoma xenograft model (compound IIII)

As a result of observation of general symptoms, no abnormality was observed in the result of death or observation of general symptoms. As a result of the body weight measurement, the body weight of the test substance 50 mg / kg / day group (G3) was significantly lower than that of the control group (G1) at 14 and 21 days after the test substance administration, And was significantly lower than that of the vehicle control group.

Tumor size measurements did not show any significant difference between all test groups, but there was a tendency to inhibit the increase in tumor size in a dose-related manner in the test substance-administered group.

In conclusion, when the test substance was administered intravenously to the malignant melanoma xenograft model using nude mouse twice / week for 3 weeks, the body weight of the test substance-administered group tended to be significantly lower or lower than that of the control group, , But there was a tendency to inhibit the increase in tumor size in a dose - correlated manner. Therefore, it was confirmed that the compound of the present invention was able to inhibit tumor growth not only in cell stage but also in animal test, and thus it was confirmed that the compound has a therapeutic and preventive effect on tumor.

Claims (10)

A compound represented by the following formula (S), a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof:

[Chemical Formula S]
In the above formula (S)
A is

As a result,
When X is O, and R ¹ is an alkoxy or hydroxy C ₁ ~ C _4,
When X is S, and R ¹ is an alkoxy or hydroxy C ₁ ~ C _4,
B is

or

As a result,
n = 1 or 2,
R ² , R ³ and R ⁴ are each independently hydrogen, tert-butyloxycarbonyl, C ₁ -C ₄ linear or branched alkyl, unsubstituted or substituted with one or more substituents, C ₅ -C ₁₂ aryl, C ₅ ~ C ₁₂ heteroaryl, or C ₅ ~ C _12, and heterocycloalkyl,
R ⁵ is X 'or X'-Y, wherein each of X' and Y is independently selected from the group consisting of C ₅ -C ₁₂ aryl, C ₅ -C ₁₂ heteroaryl, C ₅ -C ₁₂ Aryl C ₁ -C ₄ alkyl, C ₅ -C ₁₂ heteroaryl C ₁ -C ₄ alkyl, C ₅ -C ₁₂ cycloalkyl or C ₅ -C ₁₂ heterocycloalkyl,
Wherein the substituents are halogen, C ₁ ~ C ₄ linear or branched alkyl, C ₁ ~ C ₄ alkoxy, trifluoromethyl, hydroxy, amine, C ₁ ~ C ₄ alkyl amine, nitro, C ₁ ~ C ₄ alkyl Amide, and acetyl. The method according to claim 1,
When X is O, R &lt; ¹ &gt; is methoxy or hydroxy,
R ² , R ³ and R ⁴ are each independently hydrogen, tert-butyloxycarbonyl, ethyl, methyl or benzyl,
R &lt; ⁵ &gt; is phenyl, unsubstituted or substituted with one or more substituents,
Wherein said substituent is selected from the group consisting of methyl, ethyl, fluoro, trifluoromethyl, hydroxy, methoxy, nitro, and acetyl; solvates, stereoisomers or pharmaceutically acceptable salts thereof. The compound according to claim 1, wherein the compound represented by the formula (S) is a compound represented by any one of the following formulas (I) to (IV), a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof:

[Formula I] ;

[Formula II] ;

[Formula III] ; And

[Formula IV]. The compound according to claim 1, wherein the compound represented by the formula (S)
Yl) pyrimidin-2-yl) amino) ethyl) -5-methyl-2-oxo- -1,2,5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -1H-pyrazolo [3,4-d] pyrimidin- -1,2,5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -2,3-dimethyl-lH-pyrrolo [2,3- -1,2,5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydro- , 5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -4, 4-dihydro- - dimethyl-1,2,5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -6-methyl (2-oxo- -1,2,6-thiadiazinic acid 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydro- , 6-thiadiazolidin-1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -5-methyl-2-oxo- -1,2,5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -1,2,3,4-tetrahydroisoquinoline; -1,2,5-thiadiazolidine 1,1-dioxide;
2-yl) amino) ethyl) - &lt; / RTI &gt; -1,2,5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydroxy- , 5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -4, 4-dihydroxyphenyl) imidazo [ - dimethyl-1,2,5-thiadiazolidine 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -6-methyl-2-methyl- -1,2,6-thiadiazinic acid 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -1,2,3,4-tetrahydro- -1,2,6-thiadiazin-2-carboxylate 1,1-dioxide;
Yl) pyrimidin-2-yl) amino) ethyl) -1,2-dihydroxy- , 6-thiadiazolidin-1,1-dioxide;
N- (2 - ((4- (6- (3-methoxyphenyl) imidazo [2,1-b] thiazol-5-yl) pyrimidin-2-yl) amino) ethyl) benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl (2-methyl-2- ) Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl &lt; / RTI &gt; ) Benzenesulfonamide;
2-yl) amino) ethyl) -4-methyl-2-oxo- Benzenesulfonamide;
2-yl) amino) ethyl) -4- ((4-fluoropyrimidin- Trifluoromethyl) &lt; / RTI &gt;benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl &lt; / RTI &gt; ) Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl) naphthalene-2-carboxylic acid methyl ester was used in place of N- (2- (4- (6- (3- methoxyphenyl) imidazo [ Sulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl (2-oxo-2- ) Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl (2-methyl-2- ) Benzenesulfonamide;
2-yl) amino) ethyl) -4-methyl-2-oxo- Benzenesulfonamide;
2-yl) amino) ethyl) naphthalene-2-carboxylic acid methyl ester of N- (2 - ((4- (6- (4- methoxyphenyl) imidazo [ Sulfonamide;
N- (3 - ((4- (6- (3-methoxyphenyl) imidazo [2,1-b] thiazol-5-yl) pyrimidin-2-yl) amino) propyl) benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide;
Yl) amino) propyl) -4-methylpyridin-2-yl) Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) propyl) naphthalene-2-carbaldehyde, N- (3- (4- (6- (3- methoxyphenyl) imidazo [ Sulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl &lt; / RTI &gt; ) Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl &lt; / RTI &gt; ) Benzenesulfonamide;
2-yl) amino) ethyl) -4-methyl-2-pyrrolidinone Benzenesulfonamide;
Yl) pyrimidin-2-yl) amino) ethyl (2-oxo-2- ) Benzenesulfonamide;
N- (2 - ((4- (6- (4-hydroxyphenyl) imidazo [2,1-b] thiazol-5-yl) pyrimidin-2-yl) amino) ethylbenzenesulfonamide;
N- (3 - ((4- (6- (3-hydroxyphenyl) imidazo [2,1-b] thiazol-5-yl) pyrimidin-2-yl) amino) propyl) benzenesulfonamide; And
Yl) pyrimidin-2-yl) amino) propyl] -1H-pyrazolo [3,4-d] pyrimidin- ) Benzenesulfonamide, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof. Cyclizing Formulas (1) and (2) to form a compound of Formula (3);
Reacting a compound of formula 3 with 4-chloro-2- (methylthio) pyrimidine to produce a compound of formula 4;
Oxidizing the compound of formula (4) to produce a compound of formula (5); And
Reacting a compound of formula (5) with an N-aminoethylcyclic sulfonamide derivative or an N- (2-aminoethyl) benzenesulfonamide derivative under a base to obtain a compound of formula (S) Stereoisomers or pharmaceutically acceptable salts thereof.

[Reaction Scheme 1] A pharmaceutical composition for preventing and treating tumors, comprising the compound of claim 1, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient. 7. The pharmaceutical composition according to claim 6, wherein said compound, solvate, stereoisomer or pharmaceutically acceptable salt thereof inhibits protein kinase to inhibit proliferation of tumor cells. The protein kinase of claim 7, wherein the protein kinase is selected from the group consisting of V600E RAF, B-RAF, C-RAF, Mitogen-activated protein kinases 14 (MAPK14), Fms- like tyrosine kinase 3 (FLT3), and Glycogen synthase kinase 3 beta &Lt; / RTI &gt; The method of claim 6, wherein the tumor is selected from the group consisting of lung cancer, liver cancer, esophageal cancer, stomach cancer, colon cancer, small bowel cancer, pancreatic cancer, melanoma, breast cancer, oral cancer, brain tumor, thyroid cancer, papillary cancer, Cancer, bladder cancer, testicular cancer, blood cancer, lymphoma, skin cancer, psoriasis, and fibroadenoma. A method of preventing or treating a tumor, comprising administering the pharmaceutical composition of claim 6 to a subject at risk of engulfing or engulfing the tumor, wherein said subject is not a human.

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